Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

Wu Y.; Zheng Y.Y.; Lin Q.Sheng J.

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Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

机译：Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

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© 2020 Wiley Periodicals LLCThis article describes a protocol for detecting and quantifying RNA phosphorothioate modifications in cellular RNA samples. Starting from solid-phase synthesis of phosphorothioate RNA dinucleotides, followed by purification with reversed-phase HPLC, phosphorothioate RNA dinucleotide standards are prepared for UPLC-MS and LC-MS/MS methods. RNA samples are extracted from cells using TRIzol reagent, then digested with a nuclease mixture and analyzed by mass spectrometry. UPLC-MS is employed first to identify RNA phosphorothioate modifications. An optimized LC-MS/MS method is then employed to quantify the frequency of RNA phosphorothioate modifications in a series of model cells. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Synthesis, purification, and characterization of RNA phosphorothioate dinucleotides. Basic Protocol 2: Digestion of RNA samples extracted from cells. Basic Protocol 3: Detection and quantification of RNA phosphorothioate modifications by mass spectrometry.

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《current protocols in nucleic acid chemistry》 |2020年第1期|共页
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Wu Y.; Zheng Y.Y.; Lin Q.Sheng J.;
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Department of Chemistry and The RNA Institute University at Albany State University of New York;

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正文语种英语
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LC-MS/MS; nucleic acids; RNA phosphorothioate modification; UPLC-MS;

Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

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